Donepezil hydrochloride tablets

Donepezil hydrochloride tablets (lot: 110822A) were purchased from Eisai China and prepared in a mixed suspension at 0.1 mg/ml. Aβ25‐35 (lot: Y‐0044) was obtained from Bioss Biotech. Streptavidin–biotin complex and terminal deoxynucleotidyl transferase dUTP nick‐end labeling (TUNEL) apoptosis detection kits were purchased from Boster Biotech. A real‐time reverse transcription–quantitative polymerase chain reaction (RT‐qPCR) kit was obtained from TransGen Biotech. Antibodies against Bcl‐2 Bax, and β‐actin were purchased from Bio‐Rad Laboratories. A brain stereotaxic apparatus (ZH‐LAN‐STAR, C type) was obtained from Huaibei Zhenhua Biological Equipment. An inverted microscope (S70) was purchased from Leica. A ChemiDoc XRS gel imaging and analysis system was obtained from Bio‐Rad Laboratories. An SP‐9000 immunohistochemistry kit was purchased from Beijing ZSGB Biological Engineering. PCR amplification instrument (TC‐512) was from Techen. Detection kits for malondialdehyde (MDA), superoxide dismutase (SOD), glutathione (GSH) and ACHE were purchased from Nanjing Jiancheng Bioengineering Research Institute. Rabbit anti‐GFAP antibody (lot: 16825–1‐AP) was obtained from Proteintech. Rabbit anti‐IBA1 antibody (lot: bs‐1363R) was from Bioss Biotech.

8 SAMR1 male mice were used as the normal control (Control group) and 24 SAMP8 male mice were randomly divided into three groups (n = 8 per group): AD model control group (AD group), drug group (Drug group), and Electroacupuncture group (EA group). In the Drug group, donepezil hydrochloride tablets (Eisai China Inc., H20050978) were dissolved in distilled water and delivered to mice by oral gavage at a dose of 0.65 μg/g. In the EA group, mice were immobilized in mouse bags. Baihui (GV20) and Yintang (GV29) were chose for electroacupuncture for 15min per day, with transverse puncturing at a depth of 4–5 mm using disposable sterile acupuncture needles (0.25 mm × 13 mm) (Beijing Zhongyan Taihe Medicine Company). The needles were taped and connected to the HANS- LH202 electroacupuncture device (Peking University Institute of Science Nerve and Beijing Hua Wei Industrial Development Company, Beijing, China), with the sparse wave at 2 Hz, 2 V, and 0.1 mA. The mice in the Control group, AD group, and Drug group received the same 15-min restriction as the EA group.

Ten SAMR1 male mice were placed in the control group, and 30 SAMP8 male mice were randomly divided into three experimental groups of 10 mice each: the AD model control group (AD group), drug group, and electroacupuncture group (EA group). In the drug group, donepezil hydrochloride tablets (Eisai China Inc., H20050978) were dissolved in distilled water and delivered to the mice by oral gavage at a dose of 0.65 μg/g. In the EA group, mice were immobilized in mouse bags. Baihui (GV20) and Yintang (GV29) were chosen for electroacupuncture for 15 min per day, with transverse puncturing at a depth of 4–5 mm using disposable sterile acupuncture needles (0.25 mm × 13 mm) (Beijing Zhongyan Taihe Medicine Company). The needles were taped and connected to the HANS-LH202 electroacupuncture device (Peking University Institute of Science Nerve and Beijing Hua Wei Industrial Development Company, Beijing, China), with the sparse wave at 2 Hz, 2 V, and 0.1 mA. The mice in the control group, AD group, and drug group received the same 15-minute restriction as those in the EA group.

Thirty SAMP8 mice were divided into three groups (n = 10 per group): the AD group, the MA group, and the medicine (M) group. Ten SAMR1 mice were used as the normal control (N) group.
In the MA group, the mice were immobilized in mouse bags. MA on Baihui (GV20) and Yintang (GV29) was applied for 20 min, with transverse puncturing at a depth of 2–3 mm. The disposable sterile acupuncture needles (0.25 mm × 13 mm) (Beijing Zhongyan Taihe Medicine Company, Ltd.) were used. During the MA on Baihui (GV20) and Yintang (GV29), twirling manipulation was applied every 5 min and lasted 15 s each time. Each needle was rotated bidirectionally within 90° at a speed of 180°/s. For the M group, donepezil hydrochloride tablets (Eisai China, Inc., H20050978) were crushed and dissolved in distilled water and were delivered to mice by oral gavage at a dose of 0.65 μg/g (Geerts et al., 2005 (link)). The above treatments were administered once a day for 15 consecutive days, with no treatment of the N or AD groups. The mice in the N, AD, and M groups received the same 20 min restriction as the MA group. The duration of MA and selection of the acupoints was based on findings from our previous studies (Jiang et al., 2015 (link), 2016 (link), 2018 (link); Cao et al., 2017 (link); Ding et al., 2017 (link)). The above intervention lasted throughout the Morris water maze test period.